Simultaneous DNA amplification and detection using a pH-sensing semiconductor system.
Nat Methods
; 10(7): 641-6, 2013 Jul.
Article
em En
| MEDLINE
| ID: mdl-23749303
ABSTRACT
We developed an integrated chip for real-time amplification and detection of nucleic acid using pH-sensing complementary metal-oxide semiconductor (CMOS) technology. Here we show an amplification-coupled detection method for directly measuring released hydrogen ions during nucleotide incorporation rather than relying on indirect measurements such as fluorescent dyes. This is a label-free, non-optical, real-time method for detecting and quantifying target sequences by monitoring pH signatures of native amplification chemistries. The chip has ion-sensitive field effect transistor (ISFET) sensors, temperature sensors, resistive heating, signal processing and control circuitry all integrated to create a full system-on-chip platform. We evaluated the platform using two amplification strategies PCR and isothermal amplification. Using this platform, we genotyped and discriminated unique single-nucleotide polymorphism (SNP) variants of the cytochrome P450 family from crude human saliva. We anticipate this semiconductor technology will enable the creation of devices for cost-effective, portable and scalable real-time nucleic acid analysis.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Semicondutores
/
Processamento de Sinais Assistido por Computador
/
Análise de Sequência de DNA
/
Técnicas de Amplificação de Ácido Nucleico
/
Concentração de Íons de Hidrogênio
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
Nat Methods
Ano de publicação:
2013
Tipo de documento:
Article